Title |
Serological assessment of neutrophil elastase activity on elastin during lung ECM remodeling
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Published in |
BMC Pulmonary Medicine, May 2015
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DOI | 10.1186/s12890-015-0048-5 |
Pubmed ID | |
Authors |
Jacob H Kristensen, Morten A Karsdal, Jannie MB Sand, Nicholas Willumsen, Claudia Diefenbach, Birte Svensson, Per Hägglund, Diana J Oersnes-Leeming |
Abstract |
During the pathological destruction of lung tissue, neutrophil elastase (NE) degrades elastin, one of the major constituents of lung parenchyma. However there are no non-invasive methods to quantify NE degradation of elastin. We selected specific elastin fragments generated by NE for antibody generation and developed an ELISA assay (EL-NE) for the quantification of NE-degraded elastin. Monoclonal antibodies were developed against 10 NE-specific cleavage sites on elastin. One EL-NE assay was tested for analyte stability, linearity and intra- and inter-assay variation. The NE specificity was demonstrated using elastin cleaved in vitro with matrix metalloproteinases (MMPs), cathepsin G (CatG), NE and intact elastin. Clinical relevance was assessed by measuring levels of NE-generated elastin fragments in serum of patients diagnosed with idiopathic pulmonary fibrosis (IPF, n = 10) or lung cancer (n = 40). Analyte recovery of EL-NE for human serum was between 85% and 104%, the analyte was stable for four freeze/thaw cycles and after 24 h storage at 4°C. EL-NE was specific for NE-degraded elastin. Levels of NE-generated elastin fragments for elastin incubated in the presence of NE were 900% to 4700% higher than those seen with CatG or MMP incubation or in intact elastin. Serum levels of NE-generated elastin fragments were significantly increased in patients with IPF (137%, p = 0.002) and in patients with lung cancer (510%, p < 0.001) compared with age- and sex-matched controls. The EL-NE assay was specific for NE-degraded elastin. The EL-NE assay was able to specifically quantify NE-degraded elastin in serum. Serum levels of NE-degraded elastin might be used to detect excessive lung tissue degradation in lung cancer and IPF. |
Mendeley readers
Geographical breakdown
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Demographic breakdown
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Student > Ph. D. Student | 18 | 20% |
Student > Master | 12 | 14% |
Other | 6 | 7% |
Student > Bachelor | 5 | 6% |
Other | 14 | 16% |
Unknown | 12 | 14% |
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Other | 7 | 8% |
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